Abstract:

Total RNA isolation from rice seeds plays an important role in the molecular genetic basic experiments of rice. However, it is difficult to obtain high-quality rice seed RNA, which hinders the construction of cDNA library, RNA Seq and gene expression analysis. In order to explore the relevant impact factors of total RNA isolation from rice seeds and find the appropriate isolation methods, this study compared three different isolation methods, including Trizol method, GeneMark Kit Method and Adlaid kit method, different consumable treatment methods, grinding methods and other different factors to improve the effect of total RNA isolation from rice seeds. Different tests results showed that the Aidlad Polysaccharide Extraction Kit isolated total RNA from rice seeds with complete 28S, 18S bands, low degradation and high purity. Among the different consumables treatment methods, the total RNA isolated by the proteinase K-treated consumables degraded less than the other two methods. Besides, the 28S and 18S bands were clearer and more complete, and the total RNA had the highest quality and the best effect. In the grinding methods, the liquid nitrogen freeze-grinding method could inhibit the degradation of RNA by RNase effectively. In this study, the analysis and improvement of the relative factors of rice seed RNA isolation process lays a certain technical foundation for promoting the molecular experiment of rice seed as the research object.

Key words: rice seeds, RNA isolation, integrity, isolation methods, lab consumables processing